The instant invention is concerned with benzopyrancarboxylic acids and related heterocyclic compounds and pharmaceutically acceptable salts and prodrugs thereof which are useful as therapeutic compounds, particularly in the treatment and prevention of Type 2 diabetes mellitus, often referred to as non-insulin dependent diabetes (NIDDM), of conditions that are often associated with this disease, and of lipid disorders.
Diabetes refers to a disease process derived from multiple causative factors and characterized by elevated levels of plasma glucose or hyperglycemia in the fasting state or after administration of glucose during an oral glucose tolerance test. Persistent or uncontrolled hyperglycemia is associated with increased and premature morbidity and mortality. Often abnormal glucose homeostasis is associated both directly and indirectly with alterations of the lipid, lipoprotein and apolipoprotein metabolism and other metabolic and hemodynamic disease. Therefore patients with Type 2 diabetes mellitus are at especially increased risk of macrovascular and microvascular complications, including coronary heart disease, stroke, peripheral vascular disease, hypertension, nephropathy, neuropathy, and retinopathy. Therefore, therapeutical control of glucose homeostasis, lipid metabolism and hypertension are critically important in the clinical management and treatment of diabetes mellitus.
There are two generally recognized forms of diabetes. In type 1 diabetes, or insulin-dependent diabetes mellitus (IDDM), patients produce little or no insulin, the hormone which regulates glucose utilization. In type 2 diabetes, or noninsulin dependent diabetes mellitus (NIDDM), patients often have plasma insulin levels that are the same or even elevated compared to nondiabetic subjects; however, these patients have developed a resistance to the insulin stimulating effect on glucose and lipid metabolism in the main insulin-sensitive tissues, which are muscle, liver and adipose tissues, and the plasma insulin levels, while elevated, are insufficient to overcome the pronounced insulin resistance.
Insulin resistance is not primarily due to a diminished number of insulin receptors but to a post-insulin receptor binding defect that is not yet understood. This resistance to insulin responsiveness results in insufficient insulin activation of glucose uptake, oxidation and storage in muscle and inadequate insulin repression of lipolysis in adipose tissue and of glucose production and secretion in the liver.
The available treatments for type 2 diabetes, which have not changed substantially in many years, have recognized limitations. While physical exercise and reductions in dietary intake of calories will dramatically improve the diabetic condition, compliance with this treatment is very poor because of well-entrenched sedentary lifestyles and excess food consumption, especially of foods containing high amounts of saturated fat. Increasing the plasma level of insulin by administration of sulfonylureas (e.g. tolbutamide and glipizide), which stimulate the pancreatic xcex2-cells to secrete more insulin, and/or by injection of insulin after the response to sulfonylureas fails, will result in high enough insulin concentrations to stimulate the very insulin-resistant tissues. However, dangerously low levels of plasma glucose can result from these last two treatments, and increasing insulin resistance due to the even higher plasma insulin levels can occur. The biguanides increase insulin sensitivity resulting in some correction of hyperglycemia. However, the two biguanides, phenformin and metformin, can induce lactic acidosis and nausea/diarrhea, respectively.
The glitazones (i.e. 5-benzylthiazolidine-2,4-diones) are a more recently described class of compounds with potential for a novel mode of action in ameliorating many symptoms of type 2 diabetes. These agents substantially increase insulin sensitivity in muscle, liver and adipose tissue in several animal models of type 2 diabetes resulting in partial or complete correction of the elevated plasma levels of glucose without occurrence of hypoglycemia. For a review, see Willson, T. M. et al., J. Med. Chem. 43(4) 527-550, (2000).
Disorders of lipid metabolism or dyslipidemias include various conditions characterized by abnormal concentrations of one or more lipids (i.e. cholesterol and triglycerides), and/or apolipoproteins (i.e., apolipoproteins A, B, C and E), and/or lipoproteins (i.e., the macromolecular complexes formed by the lipid and the apolipoprotein that allow lipids to circulate in blood, such as Low Density Lipoproteins (LDL), Very Low Density Lipoproteins (VLDL) and Intermediate Density Lipoproteins (IDL). Cholesterol is mostly carried in Low Density Lipoproteins (LDL), and this component is commonly known as the xe2x80x9cbadxe2x80x9d cholesterol because it has been shown that elevations in IDL-cholesterol correlate closely to the risk of coronary heart disease. A smaller component of cholesterol is carried in the High Density Lipoproteins (HDL) and is commonly known as the xe2x80x9cgoodxe2x80x9d cholesterol. In fact, it is known that the primary function of HDL is to accept cholesterol deposited in the arterial wall and to transport it back to the liver for disposal through the intestine. Although it is desirable to lower elevated levels of LDL cholesterol, it is also desirable to increase levels of HDL cholesterol. Generally, it has been found that increased levels of HDL are associated with lower risk for coronary heart disease (CHD). See, for example, Gordon, et al., Am. J. Med., 62, 707-714 (1977); Stampfer, et al., N. England J. Med., 325, 373-381 (1991); and Kannel, et al., Ann. Internal Med., 90, 85-91 (1979). An example of an HDL raising agent is nicotinic acid, a drug with limited utility because doses that achieve HDL raising are associated with undesirable effects, such as flushing.
Dyslipidemias were originally classified by Fredrickson according to the combination of alterations mentioned above. The Fredrickson classification includes 6 phenotypes (i.e., I, IIa, IIb, III, IV and V) with the most common being the isolated hypercholesterolemia (or type IIa) which is usually accompained by elevated concentrations of total and LDL cholesterol. The initial treatment for hypercholesterolemia is often to modify the diet to one low in fat and cholesterol, coupled with appropriate physical exercise, followed by drug therapy when IDL-lowering goals are not met by diet and exercise alone
A second common form of dyslipidemia is the mixed or combined hyperlipidemia or type IIb and III of the Fredrickson classification. This dyslipidemia is often prevalent in patients with type 2 diabetes, obesity and the metabolic syndrome. In this dyslipidemia there are modest elevations of LDL-cholesterol, accompanied by more pronounced elevations of small dense LDL-cholesterol particles, VIDL and/or IDL (i.e., triglyceride rich lipoproteins), and total triglycerides. In addition, concentrations of HDL are often low.
Peroxisome proliferators are a structurally diverse group of compounds that when administered to rodents elicit dramatic increases in the size and number of hepatic and renal peroxisomes, as well as concomitant increases in the capacity of peroxisomes to metabolize fatty acids via increased expression of the enzymes of the beta-oxidation cycle. Compounds of this group include but are not limited to the fibrate class of lipid modulating drugs, herbicides and phthalate plasticizers. Peroxisome proliferation is also triggered by dietary or physiological factors such as a high-fat diet and cold acclimatization.
Three sub-types of peroxisome proliferator activated receptor (PPAR) have been discovered and described; they are peroxisome proliferator activated receptor alpha (PPARxcex1), peroxisome proliferator activated receptor gamma (PPARxcex3) and peroxisome proliferator activated receptor delta (PPARxcex4). Identification of PPARxcex1, a member of the nuclear hormone receptor superfamily activated by peroxisome proliferators, has facilitated analysis of the mechanism by which peroxisome proliferators exert their pleiotropic effects. PPARxcex1 is activated by a number of medium and long-chain fatty acids, and it is involved in stimulating xcex2-oxidation of fatty acids. PPARxcex1 is also associated with the activity of fibrates and fatty acids in rodents and humans. Fibric acid derivatives such as clofibrate, fenofibrate, bezafibrate, ciprofibrate, beclofibrate and etofibrate, as well as gemfibrozil, each of which are PPARxcex1 ligands and/or activators, produce a substantial reduction in plasma triglycerides as well as some increase in HDL. The effects on LDL cholesterol are inconsistent and might depend upon the compound and/or the dyslipidemic phenotype. For these reasons, this class of compounds has been primarily used to treat hypertriglyceridemia (i.e., Fredrickson Type IV and V) and/or mixed hyperlipidemia.
The PPARxcex3 receptor subtypes are involved in activating the program of adipocyte differentiation and are not involved in stimulating peroxisome proliferation in the liver. There are two known protein isoforms of PPARxcex3: PPARxcex31 and PPARxcex32 which differ only in that PPARxcex32 contains an additional 28 amino acids present at the amino terminus. The DNA sequences for the human isotypes are described in Elbrecht, et al., BBRC 224;431-437 (1996). In mice, PPARxcex32 is expressed specifically in fat cells. Tontonoz et al., Cell 79: 1147-1156 (1994) provide evidence to show that one physiological role of PPARxcex3 is to induce adipocyte differentiation. As with other members of the nuclear hormone receptor superfamily, PPARxcex32 regulates the expression of genes through interaction with other proteins and binding to hormone response elements, for example in the 5xe2x80x2 flanking regions of responsive genes. An example of a PPARxcex32 responsive gene is the tissue-specific adipocyte P2 gene. Although peroxisome proliferators, including the fibrates and fatty acids, activate the transcriptional activity of PPAR""s, only prostaglandin J2 derivatives have been identified as potential natural ligands of the PPARxcex3 subtype, which also binds thiazolidinedione antidiabetic agents with high affinity.
The human nuclear receptor gene PPARxcex4 (hPPARxcex4) has been cloned from a human osteosarcoma cell cDNA library and is fully described in A. Schmidt et al., Molecular Endocrinology, 6:1634-1641 (1992). It should be noted that PPARxcex4 is also referred to in the literature as PPARxcex2 and as NUC1, and each of these names refers to the same receptor; in Schmidt et al. the receptor is referred to as NUC1.
In WO96/01430, a human PPAR subtype, hNUC1B, is disclosed. The amino acid sequence of hNUC1B differs from human PPARxcex4 (referred to therein as hNUC1) by one amino acid, i.e., alanine at position 292. Based on in vivo experiments described therein, the authors suggest that hNUC1B protein represses hPPARxcex1 and thyroid hormone receptor protein activity.
It has been disclosed in WO97/28149 that agonists of PPARxcex4 are useful in raising HDL plasma levels. WO97/27857, 97/28115, 97/28137 and 97/27847 disclose compounds that are useful as antidiabetic, antiobesity, anti-atherosclerosis and antihyperlipidemic agents, and which may exert their effect through activation of PPARs.
It is generally believed that glitazones exert their effects by binding to the peroxisome proliferator activated receptor (PPAR) family of receptors, controlling certain transcription elements having to do with the biological entities listed above. See Hulin et al., Current Pharm. Design (1996) 2, 85-102.
A number of glitazones that are PPAR agonists have been approved for use in the treatment of diabetes. These include troglitazone, rosiglitazone and pioglitazone, all of which are primarily or exclusively PPARxcex3 agonists. Many of the newer PPAR agonists that are currently under development or are in clinical trials have dual PPARxcex1 and xcex3 activity. These are expected to improve both insulin sensitivity and the lipid profile in patients having NIDDM.
Although glitazones are beneficial in the treatment of NIDDM, there have been some serious adverse events associated with the use of the compounds. The most serious of these has been liver toxicity, which has resulted in a number of deaths. The most serious problems have occurred using troglitazone, which was recently withdrawn from the U.S. market due to these concerns about toxicity. Because of the problems that have occurred with the glitazones, researchers in a number of laboratories have been investigating classes of PPAR agonists that are not glitazones and do not contain 1,3-thiazolidinedione moieties.
Compounds that are not glitazones but are agonists of PPAR sub-types are expected to be useful in the treatment of diabetes and associated conditions. PPARxcex1 agonists should improve the lipid profile and alleviate dyslipidemias by reducing elevated LDL levels and elevated triglyceride levels and/or increasing HDL levels. PPARxcex3 agonists should improve insulin sensitivity, reducing the need for insulin injections in patients with NTDDM. The role of PPARxcex4 is less well defined.
The class of compounds described herein is novel. Structurally similar kinds of compounds have been synthesized and invesigated for other uses, particularly leukotriene B4 antagonism. See for example, ______.
The class of compounds described herein is a new class of PPAR agonists that do not contain a 1,3-thiazolidinedione moiety and therefore are not glitazones. The class of compounds includes compounds that are primarily PPARxcex1 agonists, compounds that are primarily PPARxcex3 agonists, and compounds that are mixed PPARxcex1/xcex3 agonists. The clinical effects are expected to vary depending on the balance in agonism of the PPAR-subtypes. These compounds are useful in the treatment, control and/or prevention of diabetes, hyperglycemia, mixed or diabetic dyslipidemia, and other lipid disorders (including isolated hypercholesterolemia as manifested by elevations in LDL-C and/or non-HDL-C and/or hyperapoBliproteinemia, hypertriglyceridemia and/or increase in triglyceride-rich-lipoproteins, and low HDL cholesterol concentrations), atherosclerosis, obesity, vascular restenosis, inflammatory conditions, neoplastic conditions, and other PPARxcex1 and/or xcex3 mediated diseases, disorders and conditions.
The present invention provides compounds having the structure of Formula I, including pharmaceutically acceptable salts and prodrugs of these compounds: 
In the compounds of Formula I:
Z is selected from the group consisting of CH2 and Cxe2x95x90O;
R1 is selected from H, xe2x80x94OH, C1-7alkyl, C2-7alkenyl, C2-7alkynyl, xe2x80x94OC1-3alkyl, xe2x80x94OC2-3alkenyl, xe2x80x94OC2-3alkynyl, F, Br, Cl, and Ar, wherein alkyl, alkenyl, alkynyl, xe2x80x94Oalkyl, xe2x80x94Oalkenyl and xe2x80x94Oalkynyl are linear or branched and are optionally substituted with (a) 1-7 halogen atoms and/or (b) 1-3 groups independently selected from (i) xe2x80x94OC1-3alkyl, which is optionally substituted with 1-5 halogen atoms, and (ii) phenyl, which is optionally substituted with 1-3 groups independently selected from halogen, C1-5alkyl and xe2x80x94OC1-3alkyl, said C1-5alkyl and xe2x80x94OC1-3alkyl being linear or branched and optionally substituted with 1-5 halogens; or alternatively,
R1 is a group xe2x80x94CR11R12xe2x80x94 which bridges between the carbon to which R1 is attached in FIG. I and the adjacent carbon on the heterocyclic ring, yielding a cyclopropane ring;
R11 and R12 are independently selected from the group consisting of hydrogen, halogen, C1-5alkyl, C2-5alkenyl, C2-5alkynyl, xe2x80x94OC1-3alkyl, xe2x80x94OC2-3alkenyl, xe2x80x94OC2-3alkynyl, xe2x80x94CO2H, xe2x80x94CO2C1-5alkyl, xe2x80x94CO2C2-5alkenyl, xe2x80x94CO2C2-5alkynyl, and phenyl, where alkyl, alkenyl, alkynyl, xe2x80x94Oalkyl, xe2x80x94Oalkenyl, xe2x80x94Oalkynyl xe2x80x94CO2alkyl, xe2x80x94CO2alkenyl, and xe2x80x94CO2alkynyl are linear or branched and are optionally substituted with (a) 1-5 halogens and/or (b) 1-3 substituents independently selected from xe2x80x94OCH3 and xe2x80x94OCF3, and where phenyl is optionally substituted with 1-3 groups independently selected from halogen, C1-5alkyl, and xe2x80x94OC1-3alkyl, wherein C1-5alkyl and xe2x80x94OC1-3alkyl are linear or branched and are optionally substituted with 1-5 halogens;
Ar is selected from the group consisting of Aryl, Hetcyc, Hetaryl, and Benzoheterocycle, where Aryl, Hetcyc, Hetaryl, and Benzoheterocycle are in each instance optionally substituted with 1-5 substituents independently selected from (a) halogen, (b) C1-5alkyl, (c) C2-5alkenyl, (d) C2-5alkynyl, (e) xe2x80x94OC1-5alkyl, (f) xe2x80x94OC2-5alkenyl, (g) xe2x80x94OC2-5alkynyl, (h) xe2x80x94SOxC1-5alkyl, (i) xe2x80x94SOxNRaRb, (j) xe2x80x94SOxphenyl, (k) xe2x80x94C(O)C1-3alkyl, and (l) xe2x80x94C(O)NRaRb, wherein in each instance, each alkyl, alkenyl and alkynyl is linear or branched and is optionally substituted with (a) 1-5 halogen atoms and/or (b) 1-2 groups independently selected from xe2x80x94OC1-3alkyl, which is linear or branched and is optionally substituted with 1-5 halogens, and where phenyl is optionally substituted with 1-3 substituents independently selected from halogen, C1-3alkyl, and C1-3alkoxy, wherein C1-3alkyl and C1-3alkoxy are linear or branched and are optionally substituted with 1-5 halogens, and wherein Hetcyc and Benzoheterocycle may each optionally have a C3-6-spiro-cycloalkyl substituent on the ring on a carbon atom that can have gem-disubstitution, wherein the spiro-cycloalkyl group is optionally substituted with 1-2 groups independently selected from methyl, trifluoromethyl, methoxy, trifluoromethoxy and halogen;
x is selected from 0, 1 and 2;
Aryl is a carbocyclic 6-10 membered monocyclic or bicyclic aromatic ring system;
Hetcyc is a 5- or 6-membered saturated or partly saturated monocyclic heterocycle having 1-4 heteroatoms independently selected from N, S and O in the perimeter of the ring, wherein N may optionally be NRa and S may optionally be SO or SO2;
Hetaryl is a 5- or 6-membered heteroaromatic ring having 1-4 heteroatoms independently selected from O, S, and N in the perimeter of the ring, where N may optionally be NRa, and S may optionally be SO or SO2;
Benzoheterocycle comprises a 5 or 6-membered heterocyclic ring which may be saturated, partly unsaturated or aromatic, and a benzene ring, wherein said heterocyclic ring and said benzene ring are fused together, wherein said heterocyclic ring comprises 1-3 heteroatoms independently selected from O, S, and N in the perimeter of the ring, where N may optionally be NRa, and S may optionally be SO or SO2;
Ra and Rb are independently selected from the group consisting of H, C1-5alkyl, C2-5alkenyl, C2-5alkynyl, xe2x80x94C(O)C1-5alkyl, xe2x80x94C(O)C2-5alkenyl, xe2x80x94C(O)C2-5alkynyl, SOxC1-5alkyl, SOxphenyl, SOxNRdRe, xe2x80x94C(O)NRdRe, halogen, and phenyl, wherein in all instances, alkyl, alkenyl, and alkynyl are linear or branched and are optionally substituted with (a) 1-5 halogen atoms and/or (b) 1-3 groups independently selected from xe2x80x94OCH3, xe2x80x94OCF3 and phenyl, wherein phenyl in all occurrences is optionally substituted with 1-3 substituents independently selected from halogen, C1-3alkyl, and C1-3alkoxy, said C1-3alkyl and C1-3alkoxy being linear or branched and optionally substituted with 1-5 halogens;
Rd and Re are independently selected from H, C1-5alkyl, C2-5alkenyl, C2-5alkynyl, and phenyl, wherein said alkyl, alkenyl, and alkynyl are linear or branched and are optionally substituted with (a) 1-5 halogen atoms and/or (b) 1-3 groups independently selected from xe2x80x94OCH3, xe2x80x94OCF3 and phenyl, wherein phenyl in all occurrences is optionally substituted with 1-3 substituents independently selected from halogen, C1-3alkyl, and C1-3alkoxy, said C1-3alkyl and C1-3alkoxy being linear or branched and optionally substituted with 1-5 halogens;
X and Y are independently selected from the group consisting of O, S, SO, SO2, NRa and CH2;
n is an integer from 1-6;
R2, R3, R5, R6, R7, R8, R9 and R10 are independently selected from H, halogen, C1-7alkyl, C2-7alkenyl, C2-7alkynyl, xe2x80x94OH, xe2x80x94OC1-5alkyl, xe2x80x94OC2-5alkenyl, xe2x80x94OC2-5alkynyl, xe2x80x94C(O)C1-5alkyl, xe2x80x94C(O)C2-5 alkenyl, xe2x80x94C(O)C2-5alkynyl, xe2x80x94C(O)OC1-5alkyl, xe2x80x94C(O)OC2-5alkenyl, xe2x80x94C(O)OC2-5alkynyl, xe2x80x94OC(O)C1-5alkyl, xe2x80x94OC(O)C2-5alkenyl, xe2x80x94OC(O)C2-5alkynyl, Ar, xe2x80x94OAr, xe2x80x94C(O)Ar, xe2x80x94C(O)OAr, xe2x80x94OC(O)Ar, C3-8Cycloalkyl, xe2x80x94OC3-8Cycloalkyl, xe2x80x94SOxC1-5alkyl, xe2x80x94SOxNRaRb, xe2x80x94SOxAr, and xe2x80x94C(O)NRaRb, wherein in each instance, each alkyl, alkenyl, and alkynyl is linear or branched and is optionally substituted with (a) 1-5 halogen atoms and/or (b) 1-2 groups independently selected from xe2x80x94OC1-3alkyl groups which are linear or branched and are optionally substituted with 1-5 halogens and/or (c) 1 group Ar or C3-6Cycloalkyl; and
R4 is OAr.
In the description above and elsewhere, including the claims, when something is described as being xe2x80x9coptional,xe2x80x9d such as 1 or more substituents or compounds from a list of substituents or compounds, one of the options is that the substituent or compound may be absent.
These compounds are effective in lowering glucose, lipids, and insulin in diabetic animals. The compounds are expected to be efficacious in the treatment, control and/or prevention of non-insulin dependent diabetes mellitus (NIDDM) in humans and in the treatment, control, and/or prevention of conditions associated with NIDDM, including hyperlipidemia, dyslipidemia, obesity, hypercholesterolemia, hypertrigyceridemia, atherosclerosis, vascular restenosis, inflammatory conditions, neoplastic conditions, and other PPARxcex1 and/or xcex3 mediated diseases, disorders and conditions.
The invention has numerous embodiments. Several groups of compounds are described below:
One group of compounds of formula I includes compounds in which X and Y are each O or S. Another group of compounds includes those in which X and Y are O.
One embodiment includes all compounds where Z is CH2. Another embodiment includes those compounds in which Z is Cxe2x95x90O.
Another embodiment includes those compounds in which n is 3 or 4.
A preferred group of compounds includes those compounds in which R1 is selected from Cl, Br, F and C1-4 alkyl, where C1-4alkyl is linear or branched and is optionally substituted with (a) 1-3 halogens independently selected from F and Cl, (b) 1 phenyl which is optionally substituted with 1-3 halogens, or (c) a mixture thereof.
Another preferred group of compounds of formula I includes those compounds in which R2 is selected from the group consisting of Cl, Br, F and C1-4alkyl, where C1-4alkyl is linear or branched and is optionally substituted with 1-3 halogens.
In another group of compounds having formula I, group xe2x80x94Xxe2x80x94 is attached to the benzopyran ring at the 6-position of the benzopyran ring. In a different group of compounds having formula I, the group xe2x80x94Xxe2x80x94 is attached to the benzopyran ring at the 7-position of the benzopyran ring.
In other groups of compounds having formula I, R1 is selected from C1-4alkyl, Cl and F, where alkyl is linear or branched and is optionally substituted with 1-5 F.
A preferred group of compounds of formula I comprises compounds in which Ar is phenyl, which is optionally substituted with 1-4 groups independently selected from Cl, F, C1-5alkyl, xe2x80x94OCH3, xe2x80x94OCF3, xe2x80x94SOxC1-5alkyl, xe2x80x94SOxNRaRb, xe2x80x94SOxphenyl, xe2x80x94C(O)C1-3alkyl, and xe2x80x94C(O)NRaRb, where phenyl of xe2x80x94SOxphenyl is optionally substituted with 1-3 substituents independently selected from halogen , CH3, CF3, xe2x80x94OCF3, and xe2x80x94OCH3, and where alkyl in all occurrences is linear or branched and is optionally substituted with 1-5 halogens.
In another preferred subset of compounds having Formula I, R1 and R2 are each selected from C1-4alkyl, Cl and F; n is 2-4; X and Y are O; Z is CH2; and R3, R5, R6, R7, R8, R9 and R10 are each selected from H, Cl, F, CH3, and CF3. Any alkyl in this group of compounds is linear or branched and is optionally substituted with 1-5 F. In a selected group of compounds, Z is CH2. In most compounds, R3, R5, R6, R7, R8, R9 and R10 are preferably H.
Another embodiment includes compounds having formula I as previously described, where R3, R5, R6, R7, R8, R9, and R10 are H; R2 is Cl or F; and R1 is C1-4alkyl, Cl or F, where C1-4alkyl is linear or branched and is optionally substituted with 1-5 F.
In another embodiment, R3, R5 and R6 are all H.
Another subset comprises compounds in which Ra and Rb are independently selected from H, C1-5alkyl, xe2x80x94C(O)C1-5alkyl, xe2x80x94S(O)xC1-5alkyl, xe2x80x94S(O)xphenyl, and phenyl. In these compounds, each alkyl group is linear or branched and is optionally substituted with 1-5 halogen atoms. Each phenyl is optionally substituted with 1-3 substituents independently selected from halogen, C1-3alkyl, and C1-3alkoxy, and these C1-3alkyl and C1-3alkoxy substituents are linear or branched and are optionally substituted with 1-5 halogens.
Another group of compounds is defined as those compounds having Formula I in which R1 is neither H or xe2x80x94CR11R12xe2x80x94, and no more than one of R2, R6, and R10 is alkyl.
In other preferred compounds having Formula I, R2 is Cl, Br or F, and R10 is not an alkyl group of any length.
Specific examples of compounds of this invention are provided as Examples 1-29, listed by name below. Their structures are illustrated in the Table immediately before the Examples.
The following compounds, including pharmaceutically acceptable salts and prodrugs of these compounds, are specific embodiments of this invention:
Example 1: 7-(3-(2-Propyl-4-phenoxyphenoxy)propoxy)-chromane-2-carboxylic acid;
Example 2: 7-(3-(2-Propyl-4-phenoxphenoxy)propoxy)-2-propylchromane-2-carboxylic acid;
Example 3: 7-(4-(2-Propyl-4-phenoxyphenoxy)butoxy)-chromane-2-carboxylic acid;
Example 4: 7-(4-(2-Propyl-4-phenoxyphenoxy)butoxy)-2-methylchromane-2-carboxylic acid;
Example 5: 7-(3-(2-Propyl-4-(4-chlorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 6: 7-(3-(2-Propyl-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 7: 7-(3-(2-Propyl-4-(4-methoxyphenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 8: 7-(3-(4-Phenoxyphenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 9: 7-(3-(2-Propyl-4-(4-methylsulfonylphenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 10: 7-(3-(2-Propyl-4-(4-isobutylphenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 11: 7-(2-(2-Propyl-4-phenoxyphenoxy)ethoxy)-2-ethylchromane-2-carboxylic acid;
Example 12: 7-(2-(2-Propyl-4-(4-methylsulfonylphenoxy)phenoxy)ethoxy)-2-ethylchromane-2-carboxylic acid;
Example 13: 6-(3-(2-Propyl-4-phenoxyphenoxy)propoxy)-chromane-2-carboxylic acid;
Example 14: 6-(3-(2-Propyl-4-phenoxyphenoxy)propoxy)-2-methylchromane-2-carboxylic acid;
Example 15: 6-(4-(2-Propyl-4-phenoxyphenoxy)butoxy)-chromane-2-carboxylic acid;
Example 16: 6-(4-(2-Propyl-4-phenoxyphenoxy)butoxy)-2-methyl-chromane-2-carboxylic acid;
Example 17: 7-(3-(2-Chloro-4-phenoxyphenoxy)propoxy)-chromane-2-carboxylic acid;
Example 18: 7-(3-(2-Chloro-4-phenoxyphenoxy)propoxy)-2-methylchromane-2-carboxylic acid;
Example 19: 7-(3-(2-Chloro-4-phenoxyphenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 20: 7-(3-(2-Chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-chromane-2-carboxylic acid;
Example 21: 7-(3-(2-Chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 22: 7-(3-(2-Chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-propylchromane-2-carboxylic acid;
Example 23: 7-(3-(2-Fluoro-4-phenoxyphenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 24: 7-(3-(2-Chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-(4-fluorophenylmethyl)-chromane-2-carboxylic acid;
Example 25: (2R)-7-(3-(2-Chloro-4-phenoxyphenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 26: (2R)-7-(3-(2-Chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 27: (2S)-7-(3-(2-Chloro-4-phenoxyphenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 28: (2S)-7-(3-(2-Chloro-4-(4-fluorophenoxy)phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid;
Example 29: (2R)-7-(3-(2-Propyl-4-(4-isobutylphenoxy)phenoxy)propoxy)-2-propylchromane-2-carboxylic acid.
The invention further includes pharmaceutical compositions comprising any of the compounds described above and a pharmaceutically acceptable carrier.
The compounds as defined above are useful in treating, controlling, and preventing the following diseases, and may also be used in treating other diseases not listed below:
(1) a method for treating, controlling or preventing diabetes mellitus, and particularly non-insulin dependent diabetes mellitus, in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I;
(2) a method for treating, controlling, or preventing hyperglycemia in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I;
(3) a method for treating, controlling, or preventing lipid disorders, hyperlipidemia, or low HDL in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I;
(4) a method for treating, controlling, or preventing obesity in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I;
(5) a method for treating, controlling, or preventing hypercholesterolemia in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I;
(6) a method for treating, controlling, or preventing hypertriglyceridemia in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I;
(7) a method for treating, controlling, or preventing dyslipidemia, including low HDL cholesterol, in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I;
(8) a method for treating, controlling, or preventing atherosclerosis in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I; it is understood that the sequellae of atherosclerosis (angina, claudication, heart attack, stroke, etc.) are thereby treated; and
(9) a method for treating, controlling, or preventing cachexia in a mammalian patient in need of such treatment which comprises administering to the patient a therapeutically effective amount of a compound of Formula I.
xe2x80x9cAcxe2x80x9d is acetyl, which is CH3C(O)xe2x80x94.
xe2x80x9cAlkylxe2x80x9d, as well as other groups having the prefix xe2x80x9calkxe2x80x9d, such as alkoxy or alkanoyl, means carbon chains which may be linear or branched or combinations thereof, unless the carbon chain is defined otherwise. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, and the like.
xe2x80x9cAlkenylxe2x80x9d means carbon chains which contain at least one carbon-carbon double bond, and which may be linear or branched or combinations thereof. Examples of alkenyl include vinyl, allyl, isopropenyl, pentenyl, hexenyl, heptenyl, 1-propenyl, 2-butenyl, 2-methyl-2-butenyl, and the like.
xe2x80x9cAlkynylxe2x80x9d means carbon chains which contain at least one carbon-carbon triple bond, and which may be linear or branched or combinations thereof. Examples of alkynyl include ethynyl, propargyl, 3-methyl-1-pentynyl, 2-heptynyl and the like.
xe2x80x9cCycloalkylxe2x80x9d means saturated or partly saturated monocyclic or bicyclic carbocyclic rings, each having from 3 to 10 carbon atoms, unless otherwise defined. The term also can include a monocyclic ring fused to an aryl group or other ring system. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.
xe2x80x9cArylxe2x80x9d (and xe2x80x9carylenexe2x80x9d) means mono- or bicyclic aromatic rings containing only carbon ring atoms. Aryl groups that are substituents herein are 6-10-membered monocyclic or bicyclic ring systems, and are preferably phenyl or naphthyl. Phenyl is most preferred. The term also may describe an aryl group fused to a monocyclic cycloalkyl or monocyclic heterocyclic group. xe2x80x9cHeterocyclyl,xe2x80x9d xe2x80x9cHeterocycle,xe2x80x9d and xe2x80x9cheterocyclicxe2x80x9d means a fully or partially saturated monocyclic or polycyclic ring system containing at least one heteroatom selected from N, S and O, each of said rings having from 3 to 10 atoms, except where defined otherwise. Examples of aryl include phenyl and naphthyl, as well as the phenyl ring of indanyl, indenyl, and tetrahydronaphthyl. Examples of aryl fused to heterocyclic groups include 2,3-dihydrobenzofuranyl, dihydrobenzopyranyl, and the like. Examples of heterocycles include tetrahydrofuran, piperazine, tetrahydropyran, and morpholine.
xe2x80x9cHetarylxe2x80x9d (and heteroarylene) means a mono-, bi- or tricyclic aromatic ring containing 1-4 ring heteroatoms selected from N, O and S (including SO and SO2) in the perimeter of the ring, with each ring containing 5 to 6 atoms. Examples of heteroaryl include pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl, thienyl, pyrimidyl, pyridazinyl, pyrazinyl, benzisoxazolyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl (including S-oxide and dioxide), furo(2,3-b)pyridyl, quinolyl, indolyl, isoquinolyl, dibenzofuran and the like.
xe2x80x9cHalogenxe2x80x9d includes fluorine, chlorine, bromine and iodine. Preferred halogens are chlorine and fluorine.
The term xe2x80x9ccomposition,xe2x80x9d as in pharmaceutical composition, is intended to encompass a product comprising the active ingredient(s), and the inert ingredient(s) that make up the carrier, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions of the present invention encompass any composition made by admixing a compound of the present invention and a pharmaceutically acceptable carrier.
Compounds of Formula I contain at least one asymmetric center and may contain more than one asymmetric center. The compounds can thus occur as racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The present invention is meant to comprehend all such isomeric forms of the compounds of Formula I.
The compounds of Formula I all have an asymmetric center at the 2-position of the benzopyran ring, at the position where the carboxyl group is attached to the ring. One or both enantiomers of some of the compounds described below have been isolated. The R enantiomer has higher activity than the S enantiomer in experiments performed to date, and is therefore the preferred enantiomer. Although the S enantiomer has less activity than the R enantiomer, the S enantiomer has different selectivity, and for some of the compounds has sufficient activity that it may also be useful in the treatment of PPAR mediated diseases. For example, many of the R-enantiomers have both PPARxcex1 and PPARxcex3 activity (i.e., they are PPARxcex1/xcex3 dual agonists), whereas the S enantiomers of the same compounds are often more xcex3-selective.
Some of the compounds described herein contain olefinic double bonds, and unless specified otherwise, are meant to include both E and Z geometric isomers.
Some of the compounds described herein may exist with different points of attachment of hydrogen coupled with double bond shifts, referred to as tautomers. Such an example may be a carbonyl (e.g. a ketone) and its enol form, often known as keto-enol tautomers. The individual tautomers as well as mixtures thereof are encompassed with compounds of Formula I.
If desired, racemic mixtures of compounds of Formula I may be separated by means of classical resolution through fractional crystallization of salts formed with enantiomerically pure acids or bases. Other diasteromeric derivatives can be formed by the coupling of a racemic mixture of the compounds of Formula I to an enantiomerically pure compound. Such diastereomeric mixture may be separated by standard chromatographic methods or recrystallization protocols. These diasteromeric derivatives may then be converted to the pure enantiomers of the compounds of Formula I by cleavage of the added chiral residue. The racemic mixture of the compounds of Formula I can also be separated directly by chromatographic methods utilizing chiral stationary phases, of which many examples are known in the literature.
Alternatively, any enantiomer of a compound of the general Formula I may be obtained by stereoselective synthesis using optically pure starting materials or reagents of known configuration.
Compounds of Formula I that have more than one asymmetric center and that occur as diasteromeric mixtures can similarly be separated into individual diastereomers by standard methods, and these can be separated to individual enantiomers as described above.
The term xe2x80x9cpharmaceutically acceptable saltsxe2x80x9d refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic or organic bases and inorganic or organic acids. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts. Salts in the solid form may exist in more than one crystal structure, and may also be in the form of hydrates. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,Nxe2x80x2-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.
When the compound of the present invention is basic, salts may be organic acids. Such acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid, and the like. Particularly preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric, and tartaric acids.
It will be understood that, as used herein, references to the compounds of Formula I are meant to also include the pharmaceutically acceptable salts.
This invention also includes the active metabolites of claimed compounds. Prodrugs, which are compounds that are converted to the claimed compounds as they are being administered to a patient or after they have been administered to a patient, are also included within the scope of the claimed active compounds. A non-limiting example of a prodrug of the carboxylic acids of this invention would be an ester of the carboxylic acid group, for example a C1 to C6 ester, which may be linear or branched, or an ester which has functionality that makes it more easily hydrolyzed after administration to a patient.
Examples of prodrugs of this class of compounds may be described as compounds having the Formula Ia: 
R1,R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, Ra, Rb, Rd, Re, X, Y, Z, n, x, Ar, and other substituents are as defined previously. In the prodrugs, W is a group that is easily removed under physiological conditions during or after administration to a mammalian patient to yield a compound having Formula I, or the carboxylate anion thereof (in solution), or a pharmaceutically acceptable salt thereof.
Examples of prodrugs of Formula Ia include compounds in which W is selected from the group consisting of xe2x80x94OR13, OCH2OR13, xe2x80x94OCH(CH3)OR13, xe2x80x94OCH2OC(O)R13, xe2x80x94OCH(CH3)OC(O)R13, xe2x80x94OCH2OC(O)OR13, xe2x80x94OCH(CH3)OC(O)OR13, and xe2x80x94NR14R14, where each R13 is independently selected from C1-6 alkyl optionally substituted with one or two groups selected from xe2x80x94CO2H, xe2x80x94CONH2, xe2x80x94NH2, xe2x80x94OH, xe2x80x94OAc, xe2x80x94NHAc, and phenyl; and wherein each R14 is independently selected from H and R13. Compounds having Formula Ia, where W has the chemical structure described above, are described as prodrugs. However, regardless of whether they are active as prodrugs, yielding compounds or salts of Formula I, or whether they have a different means of exhibiting pharmaceutical activity, the compounds of Formula Ia are included in this invention. Such compounds are claimed herein, regardless of the mechanism leading to their activity.
The description of utility, pharmaceutical compositions, combination therapies, administration, dosage, and the like that are described herein are applicable to the prodrugs described above and to the compounds described previously.
Compounds of the present invention are potent agonists of various peroxisome proliferator activator receptor subtypes, particularly PPARxcex1 and/or PPARxcex3. Compounds of the present invention may be selective agonists of one receptor subtype, e.g. PPARxcex3 or PPARxcex1 agonists, or they may be agonists of more than one receptor subtypes, e.g. dual PPARxcex1/xcex3 agonists. Compounds of the present invention are useful in treating, controlling or preventing diseases, disorders or conditions, wherein the treatment is mediated by the activation of an individual PPAR subtype (xcex1 or xcex3), or a combination of PPAR subtypes (e.g. xcex1/xcex3). Thus one aspect of the present invention provides a method for the treatment, control or prevention of such diseases, disorders, or conditions in a mammal which comprises administering to such mammal a therapeutically effective amount of a compound of Formula I. The diseases, disorders or conditions for which compounds of the present invention are useful in treating, controlling or preventing include, but are not limited to, (1) diabetes mellitus, and especially non-insulin dependent diabetes mellitus (NIDDM), (2) hyperglycemia, (3) impaired glucose tolerance, (4) insulin resistance, (5) obesity, (6) lipid disorders, (7) dyslipidemia, (8) hyperlipidemia, (9) hypertriglyceridemia, (10)hypercholesterolemia, (11) low HDL levels, (12) high LDL levels, (13) atherosclerosis and its sequelae, (14) vascular restenosis, (15) irritable bowel syndrome, (16) inflammatory bowel disease, including Crohn""s disease and ulcerative colitis, (17) other inflammatory conditions, (18) pancreatitis, (19) abdominal obesity, (20) neurodegenerative disease, (21) retinopathy, (22) neoplastic conditions, (23) adipose cell tumors, (24) adipose cell carcinomas, such as liposarcoma, (25) prostate cancer and other cancers, including gastric, breast, bladder and colon cancers, (26) angiogenesis, (27) Alzheimer""s disease, (28) psoriasis, (29) acne vulgaris, (30) other skin diseases and dermatological conditions modulated by PPAR, (31) high blood pressure, (32) Syndrome X, (33) ovarian hyperandrogenism (polycystic ovarian syndrome), and other disorders where insulin resistance is a component.
Another aspect of the invention provides a method for the treatment, control, or prevention of hypercholesterolemia, atherosclerosis, low HDL levels, high LDL levels, hyperlipidemia, hypertriglyceridemia, and/or dyslipidemia, which comprises administering to a mammal in need of such treatment a therapeutically effective amount of an agonist of PPARxcex1 and/or PPARxcex3 or a PPARxcex1/xcex3 dual agonist. The PPAR agonist may be used alone or advantageously may be administered with a cholesterol biosynthesis inhibitor, including but not limited to, an HMG-CoA reductase inhibitor such as lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rivastatin, itavastatin, or ZD-4522. The PPAR agonist may also be used advantageously in combination with other lipid lowering drugs such as cholesterol absorption inhibitors (for example stanol esters, sterol glycosides such as tiqueside, and azetidinones such as ezetimibe), ACAT inhibitors (such as avasimibe), and with niacin, bile acid sequestrants, microsomal triglyceride transport inhibitors, and bile acid reuptake inhibitors. These combination treatments may also be effective for the treatment, control or prevention of one or more related conditions selected from the group consisting of hypercholesterolemia, atherosclerosis, hyperlipidemia, hypertriglyceridemia, dyslipidemia, high LDL, and low HDL.
Another aspect of the invention provides a method of treating inflammatory conditions, including inflammatory bowel disease, Crohn""s disease, and ulcerative colitis by administering an effective amount of a PPAR agonist, which may be a PPARxcex1 agonist, a PPARxcex3 agonist, or a PPARxcex1/xcex3 dual agonist. Additional inflammatory diseases that may be treated with the instant invention include gout, rheumatoid arthritis, osteoarthritis, multiple sclerosis, asthma, ARDS, psoriasis, vasculitis, ischemia/reperfusion injury, frostbite, and related diseases.
Another aspect of the invention provides a method of treating cachexia. PPARxcex1 is known to be necessary for an appropriate energy sparing response to starvation, and inappropriate metabolism and energy utilization is clearly responsible for the wasting of cachexia.
Another aspect of the invention provides a method of treating a variety of skin diseases and dermatological conditions that are modulated by PPARxcex1 and/or xcex3 agonists. These diseases and conditions include psoriasis and acne vulgaris. Examples of other skin diseases and dermatological disorders that may be treated include eczema; lupus associated skin lesions; dermatitides such as seborrheic dermatitis and solar dermatitis; keratoses such as seborrheic keratosis, senile keratosis, actinic keratosis, photo-induced keratosis, and keratosis follicularis; keloids and prophylaxis against keloid formation, warts including verruca, condyloma, or condyloma accuminatum, and human papilloma viral (HPV) infections such as venereal warts, viral warts, molluscum contagiosum, leukoplakia, lichen planus; keratitis, skin cancer such as basal cell carcinoma, cutaneous T cell lymphoma and localized benign epidermal tumors (keratoderma, epidermal naevi).
Any suitable route of administration may be employed for providing a mammal, especially a human, with an effective dose of a compound of the present invention. For example, oral, rectal, topical, parenteral, ocular, pulmonary, nasal, and the like may be employed. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, creams, ointments, aerosols, and the like. Preferably compounds of Formula I are administered orally.
The effective dosage of active ingredient employed may vary depending on the particular compound employed, the mode of administration, the condition being treated and the severity of the condition being treated. Such dosage may be ascertained readily by a person skilled in the art.
When treating or preventing diabetes mellitus and/or hyperglycemia or hypertriglyceridemia or other diseases for which compounds of Formula I are indicated, generally satisfactory results are obtained when the compounds of the present invention are administered at a daily dosage of from about 0.1 milligram to about 100 milligram per kilogram of animal body weight, preferably given as a single daily dose or in divided doses two to six times a day, or in sustained release form. For most large mammals, the total daily dosage is from about 1.0 milligrams to about 1000 milligrams, preferably from about 1 milligrams to about 50 milligrams. In the case of a 70 kg adult human, the total daily dose will generally be from about 7 milligrams to about 350 milligrams. This dosage regimen may be adjusted to provide the optimal therapeutic response.
Another aspect of the present invention provides pharmaceutical compositions which comprise a compound of Formula I and a pharmaceutically acceptable carrier. The pharmaceutical compositions of the present invention comprise a compound of Formula I or a pharmaceutically acceptable salt or prodrug thereof as an active ingredient, as well as a pharmaceutically acceptable carrier and optionally other therapeutic ingredients. The term xe2x80x9cpharmaceutically acceptable saltsxe2x80x9d refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic bases or acids and organic bases or acids.
The compositions include compositions suitable for oral, rectal, topical, parenteral (including subcutaneous, intramuscular, and intravenous), ocular (ophthalmic), pulmonary (nasal or buccal inhalation), or nasal administration, although the most suitable route in any given case will depend on the nature and severity of the conditions being treated and on the nature of the active ingredient. They may be conveniently presented in unit dosage form and prepared by any of the methods well-known in the art of pharmacy.
In practical use, the compounds of Formula I can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). In preparing the compositions for oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like in the case of oral liquid preparations, such as, for example, suspensions, elixirs and solutions; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like in the case of oral solid preparations such as, for example, powders, hard and soft capsules and tablets, with the solid oral preparations being preferred over the liquid preparations.
Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques. Such compositions and preparations should contain at least 0.1 percent of active compound. The percentage of active compound in these compositions may, of course, be varied and may conveniently be between about 2 percent to about 60 percent of the weight of the unit. The amount of active compound in such therapeutically useful compositions is such that an effective dosage will be obtained. The active compounds can also be administered intranasally as, for example, liquid drops or spray.
The tablets, pills, capsules, and the like may also contain a binder such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose or saccharin. When a dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a fatty oil.
Various other materials may be present as coatings or to modify the physical form of the dosage unit. For instance, tablets may be coated with shellac, sugar or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye and a flavoring such as cherry or orange flavor.
Compounds of formula I may also be administered parenterally. Solutions or suspensions of these active compounds can be prepared in water suitably mixed with a surfactant such as hydroxy-propylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.
Compounds of Formula I may be used in combination with other drugs that may also be useful in the treatment, prevention, suppression or amelioration of the diseases or conditions for which compounds of Formula I are useful. Such other drugs may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with a compound of Formula I. When a compound of Formula I is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and the compound of Formula I is preferred. However, the combination therapy also includes therapies in which the compound of Formula I and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compound of the present invention and the other active ingredients may be used in lower doses than when each is used singly. Accordingly, the pharmaceutical compositions of the present invention include those that contain one or more other active ingredients, in addition to a compound of Formula I.
Examples of other active ingredients that may be administered in combination with a compound of Formula I, and either administered separately or in the same pharmaceutical composition, include, but are not limited to:
(a) insulin sensitizers including (i) PPARxcex3 agonists such as the glitazones (e.g. troglitazone, pioglitazone, englitazone, MCC-555, rosiglitazone, and the like), and compounds disclosed in WO97/27857, 97/28115, 97/28137 and 97/27847; (ii) biguanides such as metformin and phenformin; (iii) protein tyrosine phosphatase-1B (PTP-1B) inhibitors, and (iv) dipeptidyl peptidase IV (DP-IV) inhibitors;
(b) insulin or insulin mimetics;
(c) sulfonylureas such as tolbutamide and glipizide, or related materials;
(d) xcex1-glucosidase inhibitors (such as acarbose);
(e) cholesterol lowering agents such as (i) EDNG-CoA reductase inhibitors (lovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin, rivastatin, itavastatin, 2D-4522 and other statins), (ii) sequestrants (cholestyramine, colestipol, and dialkylaminoalkyl derivatives of a cross-linked dextran), (iii) nicotinyl alcohol, nicotinic acid or a salt thereof, (iv) PPARxcex1 agonists such as fibric acid derivatives (clofibrate, fenofibrate and bezafibrate) or gemfibrozil (v) PPARxcex1/xcex3 dual agonists, such as KRP-297, (vi) inhibitors of cholesterol absorption, such as for example ezetimibe, (vii) acyl CoA:cholesterol acyltransferase inhibitors, such as for example avasimnibe, and (viii) anti-oxidants, such as probucol;
(f) PPARxcex4 agonists such as those disclosed in WO97/28149;
(g) antiobesity compounds (anorectics) such as fenfluramine, dexfenfluramine, phentermine, sibutramine, mazindol, orlistat, lipase inhibitors, neuropeptide Y5 inhibitors, and xcex23 adrenergic receptor agonists;
(h) an ileal bile acid transporter inhibitor; and
(i) agents intended for use in inflammatory conditions such as aspirin, non-steroidal anti-inflammatory drugs, glucocorticoids, azulfidine, and cyclo-oxygenase 2 selective inhibitors.
The above combinations include combinations of a compound of the present invention not only with one other active compound, but also with two or more other active compounds. Non-limiting examples include combinations of compounds having Formula I with two or more active compounds selected from biguanides, sulfonylureas, HMG-CoA reductase inhibitors, other PPAR agonists, PTP-1B inhibitors, DP-IV inhibitors, and anti-obesity compounds.
A) PPAR Binding Assays
For preparation of recombinant human PPARxcex3, PPARxcex4, and PPARxcex1: Human PPARxcex32, human PPARxcex4 and human PPARxcex1 were expressed as gst-fusion proteins in E. coli. The full length human cDNA for PPARxcex32 was subcloned into the pGEX-2T expression vector (Pharmacia). The full length human cDNAs for PPARxcex4 and PPARxcex1 were subcloned into the pGEX-KT expression vector (Pharmacia). E. coli containing the respective plasmids were propagated, induced, and harvested by centrifugation. The resuspended pellet was broken in a French press and debris was removed by centrifugation at 12,000xc3x97g. Recombinant human PPAR receptors were purified by affinity chromatography on glutathione sepharose. After application to the column, and one wash, receptor was eluted with glutathione. Glycerol (10%) was added to stabilize the receptor and aliquots were stored at xe2x88x9280xc2x0 C.
For binding to PPARxcex3, an aliquot of receptor was incubated in TEGM (10 mM Tris, pH 7.2, 1 mM EDTA, 10% glycerol, 7 xcexcL/100 mL xcex2-mercaptoethanol, 10 mM Na molybdate, 1 mM dithiothreitol, 5 xcexcg/mL aprotinin, 2 xcexcg/mL leupeptin, 2 xcexcg/mL benzamidine and 0.5 mM PMSF) containing 0.1% non-fat dry milk and 10 nM [3H2] AD5075, (21 Ci/mmole), xc2x1test compound as described in Berger et al (Novel peroxisome proliferator-activated receptor (PPARxcex3) and PPARxcex4 ligands produce distinct biological effects. J. Biol. Chem. (1999), 274: 6718-6725.) Assays were incubated for xcx9c16 hr at 4xc2x0 C. in a final volume of 150 xcexcL. Unbound ligand was removed by incubation with 100 xcexcL dextran/gelatin-coated charcoal, on ice, for xcx9c10 min. After centrifugation at 3000 rpm for 10 min at 4xc2x0 C., 50 xcexcL of the supernatant fraction was counted in a Topcount.
For binding to PPARxcex4, an aliquot of receptor was incubated in TEGM (10 mM Tris, pH 7.2, 1 mM EDTA, 10% glycerol, 7 xcexcL/100 mL xcex2-mercaptoethanol, 10 mM Na molybdate, 1 mM dithiothreitol, 5 xcexcg/mL aprotinin, 2 xcexcg/mL leupeptin, 2 xcexcg/mL benzamide and 0.5 mM PMSF) containing 0.1 % non-fat dry milk and 2.5 nM [3H2]L-783483, (17 Ci/mmole), xc2x1test compound as described in Berger et al (Novel peroxisome proliferator-activated receptory (PPARxcex3) and PPARxcex4 ligands produce distinct biological effects.1999 J Biol Chem 274: 6718-6725). (L-783483 is 3-chloro-4-(3-(7-propyl-3-trifluoromethyl-6-benz-[4,5]-isoxazoloxy)propylthio)phenylacetic acid, Ex. 20 in WO 97/28137). Assays were incubated for xcx9c16 hr at 4xc2x0 C. in a final volume of 150 xcexcL. Unbound ligand was removed by incubation with 100 xcexcL dextran/gelatin-coated charcoal, on ice, for xcx9c10 min. After centrifugation at 3000 rpm for 10 min at 4xc2x0 C., 50 xcexcL of the supernatant fraction was counted in a Topcount.
For binding to PPARxcex1, an aliquot of receptor was incubated in TEGM (10 mM Tris, pH 7.2, 1 mM EDTA, 10% glycerol, 7 xcexcL/100 mL xcex2-mercaptoethanol, 10 mM Na molybdate, 1 mM dithiothreitol, 5 xcexcg/mL aprotinin, 2 ag/mL leupeptin, 2 xcexcg/mL benzamide and 0.5 mM PMSF) containing 0.1% non-fat dry milk and 5.0 nM [3H2](3-(4-(3-phenyl-7-propyl-6-benz-[4,5]-isoxazoloxy)butyloxy))phenylacetic acid (34 Ci/mmole), xc2x1test compound. This is a tritium labelled variant of Ex.62 in WO 97/28137. Assays were incubated for xcx9c16 hr at 4xc2x0 C. in a final volume of 150 xcexcL. Unbound ligand was removed by incubation with 100 xcexcL dextran/gelatin-coated charcoal, on ice, for xcx9c10 min. After centrifugation at 3000 rpm for 10 min at 4xc2x0 C., 50 xcexcL of the supernatant fraction was counted in a Topcount.
B). Gal-4 hPPAR Transactivation Assays
The chimeric receptor expression constructs, pcDNA3-hPPARxcex3/GAL4, pcDNA3-hPPARxcex4/GAL4, pcDNA3-hPPARxcex1/GAL4 were prepared by inserting the yeast GAL4 transcription factor DBD adjacent to the ligand binding domains (LBDs) of hPPARxcex3, hPPARxcex4, hPPARxcex1, respectively. The reporter construct, pUAS(5X)-tk-luc was generated by inserting 5 copies of the GAL4 response element upstream of the herpes virus minimal thymidine kinase promoter and the luciferase reporter gene. pCMV-lacZ contains the galactosidase Z gene under the regulation of the cytomegalovirus promoter. COS-1 cells were seeded at 12xc3x97103 cells/well in 96 well cell culture plates in high glucose Dulbecco""s modified Eagle medium (DMEM) containing 10% charcoal stripped fetal calf serum (Gemini Bio-Products, Calabasas, Calif.), nonessential amino acids, 100 units/ml Penicillin G and 100 mg/ml Streptomycin sulfate at 37 xc2x0 C. in a humidified atmosphere of 10% CO2. After 24 h, transfections were performed with Lipofectamine (GIBCO BRL, Gaithersburg, Md.) according to the instructions of the manufacturer. Briefly, transfection mixes for each well contained 0.48 xcexcl of Lipofectamine, 0.00075 xcexcg of pcDNA3-PPAR/GALA expression vector, 0.045 xcexcg of pUAS(5X)-tk-luc reporter vector and 0.0002 xcexcg of pCMV-lacZ as an internal control for transactivation efficiency. Cells were incubated in the transfection mixture for 5 h at 37xc2x0 C. in an atmosphere of 10% CO2. The cells were then incubated for xcx9c48 h in fresh high glucose DMEM containing 5% charcoal stripped fetal calf serum, nonessential amino acids, 100 units/mi Penicillin G and 100 mg/ml Streptomycin sulfatexc2x1increasing concentrations of test compound. Since the compounds were solubilized in DMSO, control cells were incubated with equivalent concentrations of DMSO; final DMSO concentrations were xe2x89xa60.1%, a concentration which was shown not to effect transactivation activity. Cell lysates were produced using Reporter Lysis Buffer (Promega, Madison, Wis.) according to the manufacturer""s instructions. Luciferase activity in cell extracts was determined using Luciferase Assay Buffer (Promega, Madison, Wis.) in an ML3000 luminometer (Dynatech Laboratories, Chantilly, Va.). xcex2-galactosidase activity was determined using xcex2-D-galactopyranoside (Calbiochem, San Diego, Calif.).
C. In Vivo Studies
Male db/db mice (10-11 week old C57BI/KFJ, Jackson Labs, Bar Harbor, Me.) were housed 5/cage and allowed ad lib. access to ground Purina rodent chow and water. The animals, and their food, were weighed every 2 days and were dosed daily by gavage with vehicle (0.5% carboxymethylcellulose)xc2x1test compound at the indicated dose. Drug suspensions were prepared daily. Plasma glucose, and triglyceride concentrations were determined from blood obtained by tail bleeds at 3-5 day intervals during the study period. Glucose, and triglyceride, determinations were performed on a Boehringer Mannheim Hitachi 911 automatic analyzer (Boehringer Mannheim, Indianapolis, Ind.) using heparinized plasma diluted 1:6 (v/v) with normal saline. Lean animals were age-matched heterozygous mice maintained in the same manner.
The process for making the compounds of the instant invention is generally described in Scheme 1 shown below. 
The appropriately substituted benzopyran carboxylate of formula III (E is alkyl or aryl, for example, a methyl or ethyl group) may be synthesized by the coupling of compounds having formulae VII and IV, or by the coupling of compounds having formulae V and VI, where coupling is carried out in the presence of inorganic base (e.g. cesium carbonate) in DMF, or under standard Mitsunobu reaction condition (e.g. diiosopropyl azodicarboxylate and triphenyl phosphine) in dichloromethane. Lv(1) and LV(2) are leaving groups well-known in the art, and preferably are independently selected from halogen, preferably bromine, or sulfonate such as methanesulfonate or p-toluenesulfonate, or a hydroxyl group. Compounds having formulae VII and V may be commercially available, or prepared by published organic synthetic methods. The desired benzopyran carboxylic acid I may be synthesized by ester hydrolysis of the compound having formula III under aqueous basic (e. g. aq. NaOH) or acidic conditions.
Optionally, when the R1 group in formula III is hydrogen, an R1 group other than hydrogen may be introduced under standard ester enolate alkylation conditions (e.g. using sodium bis(trimethylsilyl)amide and R1xe2x80x94Lv(3) in TEE solvent at low temperature, wherein Lv(3) is a leaving group, preferably iodine or bromine, to give a compound having formula II. The desired benzopyran carboxylic acids I may be synthesized by ester hydrolysis of the compound having formula II under aqueous basic (e.g. aq. NaOH) or acidic conditions.